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Prolactine bij Teleostei: kwantitatieve opsporing met fluorchroom gekoppelde antilichamen
Van Houtte, A. (1976). Prolactine bij Teleostei: kwantitatieve opsporing met fluorchroom gekoppelde antilichamen. MSc Thesis. Katholieke Universiteit Leuven. Faculteit Wetenschappen: Leuven. 78 pp.
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| Beschikbaar in | Auteur |
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Documenttype: Doctoraat/Thesis/Eindwerk
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| Trefwoorden |
Fluids > Body fluids > Serum > Antibodies
Secretory products > Hormones
Tracer techniques
Teleostei [WoRMS]
Marien/Kust |
| Abstract |
The presence of prolactin in teleosts has been confirmed in literature in various ways. The hormone shows physiological importance especially in osmoregulation and water economy in species living in fresh water. Other roles attributed to prolactin concern reproduction,brooding habits and pigmentation dispersion, as can be indicated in some species.Extensive experimental probing of the influence of prolactin is limited to some specialized laboratories through the lack of a simple quantitative detection method.Prolactin is a protein hormone synthetized by acidophilic cells of the pars distalis of the hypophysis. Sheep prolactine is therefore antigenetically active, can be extracted and be used for antibody production and for being labelled. The hormone is assayed by means of immunochemical methods.The principle of the classical assay is based on the competitive binding of prolactin and a radioactive labelled prolactin fraction with a limited quantity of antibody receptor molecules.In our study we have looked into the possibility of augmenting the sensibility of other immunochemical methods in order to detect the prolact in contents of fish serum. Therefore we used antibodies against sheep prolactin coupled to fluorescein isothiocyanate.We tried out precipitation methods in gels. The sensibility in the homologous radial immunodiffusion method allows detection of 200 ng sheep prolactin/ml solution. In this way no prolactin in serum can be traced.In analogy with immunoradiometric methods we worked out a precipitation method in a buffer system. The prolactin antigens are incubated with an excess of fluorochrome coupled antibodies. The remaining antibodies are separated from the formed immunocomplex by means of an antigen immunoadsorbent. The fluorescence of the medium was meant to be standardized as a measure for prolactin quantity. Experiments however point out that when measuring the fluorescence of the solution, errors occur which seem theoretically impredictable. The results are not in relation to the given amount of prolactin antigenes. |
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